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Preparation of the Antibodies of Peanuts and Shrimp Allergen and Simultaneous Detemination by Piezoelectric Immunosensor

Author: NingZuo
Tutor: QianHe
School: Jiangnan University
Course: Nutrition and Food Hygiene
Keywords: Peanut 河虾 Enzyme-linked immunosorbent Piezoelectric Immunosensor Synchronous detection
CLC: R392.1
Type: Master's thesis
Year: 2009
Downloads: 101
Quote: 1
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Abstract


Peanuts, shrimp belonging to eight categories prone to food allergies, relative to other food allergies in terms of their allergy incidence, clinical symptoms are more severe, much attention in the field of public health and food safety. Existing the allergen detection method is cumbersome, unable to meet the fast, easy, online testing requirements, therefore, the research and the establishment of a convenient, safe and fast detection method has great significance. Study including peanuts, shrimp allergens isolated and purified rabbit anti-peanut, shrimp preparation of polyclonal antibody, to to explore colloidal gold can be used as a new type of immune adjuvant; piezoelectric immunosensor use of self-assembled determination peanut The the immune affinity constant of the antibody and shrimp antibody, and at the same time package is two antibodies to allergens were screening. Peanut, shrimp protein immersion by ammonium sulfate precipitation, isoelectric point precipitation, AKTA protein purification system to obtain a purified, collected after electrophoresis test. Using sodium citrate reduction method to get around 16nm colloidal gold, colloidal gold to New Zealand white rabbits were set up as a model animal for the preparation of antibodies, immune process as adjuvant and Freund's complete (incomplete) adjuvant as a control, able to find colloidal gold significantly improve the White Rabbit in vivo anti-peanut the river shrimp allergy protein antibody levels, peanuts and colloidal gold group average titer Freund's adjuvant group doubling its IC 50 34.28 ng / mL; river shrimp with colloidal gold group average titer of Freund's adjuvant group a 25% increase in its IC 50 43.95 ng / mL. The use of self-assembled piezoelectric immunosensor determination peanut antibody affinity constant of the Kα = 2.86 × 10 7 L / mol, the hybrid antibodies of peanut antigens, the Kα = 1.37 × 10 7 L / mol; the Kα of 河虾 antibodies = 5.16 × 10 7 L / mol, mixed of antibodies against 河虾 antigen-Kα = 2.57 × 10 7 L / mol . The self-assembly of piezoelectric sensor package peanut allergens in peanuts and shrimp mixed antibodies were added to a series of concentration and river shrimp allergen detection, draw the standard curve and detection sensitivity. Simultaneous synchronization screening spiked recovery experiments, high recovery, good reproducibility, peanut, shrimp allergy original. Established in this study self-assembly of the piezoelectric sensor package at the same time the two antibodies peanuts, shrimp, achieve synchronization of peanuts, shrimp allergen detection, fast, convenient and has good practical value.

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CLC: > Medicine, health > Basic Medical > Medical Immunology > Immunobiology
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